Two isoforms of the tick Ixodes ricinus digestive peptidases cathepsins L1 and L3 are expressed also in salivary glands. In order to assess their role in this tissue, the specific antibodies against these peptidases were prepared by affinity chromatography and used for their detailed immuno-localization using cutting-edge methods of electron microscopy.

The overall structure of photosynthetic pigment-protein complexes and thylakoid membranes of various photosynthetic organisms was studied using electron microscopy.

The aim of this study was to define and examin the capabilities of the low voltage transmission electron microscope LVEM 5 in the detection and identification of viruses, taking into consideration various contrasting agents, and a comparison to data from HV TEM.

I have set for a challenging study that combined experimental and theoretical approaches in an attempt to resolve a role of small aminoacids in intermolecular interactions. First, I have proposed a hypothesis that described the interaction among individual aminoacids forming D helices of D1 and D2 proteins based on molecular dynamic simulations of a simplified model representing the reaction centre of photosystem II. Stability of the putative interhelical hydrogen bond network connecting D1 and D2 proteins was investigated experimentally with dynamic force spectroscopy using atomic force microscope. The results of both methods are in a full agreement with each other and reveal the key role of D1-Gly208 aminoacid in stability and functionality of photosystem II by providing milieu for weak interactions among three contact points at the cross of D helices: D1-Gly208 (O) and D2-Cys211 (O?), D1-Ser209 (O?) and D2-Ile204 (O), D1-Ser212 (O?) and D2-Gly207 (O). Mutation of the D1-Gly208 led to the increase in probability of the binding among the aforementioned aminoacids, undesirably strengthening the overall interactions among the proteins compromising photosynthetic capacity (D1-Ser208) or disabling of autotrophic growth (D1-Val208).

Transcription has turned out to be a discontinuous process when imaged at a single cell level. This observation is referred to as transcriptional bursting or pulsing and has been detected in a variety of organisms ranging from bacteria to mammalian cells. The dynamics of transcriptional pulsing are influenced by the properties intrinsic to the transcriptional process, as well as by upstream factors: chromatin environment, signalling molecules, cell cycle stage etc. In the first part of this thesis, we focused on the regulation of transcriptional pulsing in the nucleolus. Using imaging of living cells, we detected pulsatile transcription of a transgene with nucleolar localization whose expression was mediated by RNA polymerase II. In the second part of the thesis, we investigated the relationship between chromatin decondensation and transcriptional dynamics. We used hyperosmotic medium to induce global condensation of chromatin and revealed that upon chromatin decondensation, a transient spike in transcriptional intensity occurs in induvial living cells. Next, we analysed expression of TFRC and POLR2A genes in several cell cycle stages using single molecule RNA FISH. We detected increase in both frequency and size of transcriptional pulses during a limited time window which coincided with chromatin...

This thesis is interested in new materials for lithium – ion batteries. Two different samples were investigated, one intercalation and one conversion cathode material. The theoretical part is focused to the structure of cells, their advantages and disadvantages compared to other secondary batteries. Also other materials used in batteries are described. The practical part describes the preparation of cathode materials for subsequent testing by scanning elektron microscopy and thermogravimetric analysis. In conclusions two different materials were evaluated and compared with each other.

This thesis deals with monitoring the durability of FRP reinforcement in different types of environments which cause degradation. The theoretical part of the thesis describes reinforcing fibers, matrix materials, manufacturing technology of composite reinforcements by pultrusion and durability of FRP materials in environmental degradation. In the experimental part of this thesis was carried storing samples reinforcements in alkaline environment with various temperatures. As suitable were selected temperatures of 20 °C, 40 °C and 60 °C. Subsequently are described changes in physical properties of the reinforcements and tracking optical microscope.

We present a fluid-membrane lens with two piezoelectric actuators that offer versatile, circular symmetric lens surface shaping. A wavefront-measurement-based control system ensures robustness against creeping and hysteresis effects of the piezoelectric actuators. We apply the adaptive lens to correct synthetic aberrations induced by a deformable mirror. The results suggest that the lens is able to correct spherical aberrations with standard Zernike coefficients between 0 μm and 1 μm, while operating at refractive powers up to about 4m-1. We apply the adaptive lens in a custom-built confocal microscope to allow simultaneous axial scanning and spherical aberration tuning. The confocal microscope is extended by an additional phase measurement system to include the control algorithm. To verify our approach, we use the maximum intensity and the axial FWHM of the overall confocal point spread function as figures of merit. We further discuss the ability of the adaptive lens to correct specimen-induced aberrations in a confocal microscope.

Congenital disorders of glycosylation (CDG) are rare multisystem metabolic diseases and their number has rapidly grown in recent years. The clinical manifestation includes very broad spectrum of symptoms. In most of all cases CDG are caused by mutations in genes encoding the enzymes of glycosylation pathway. Based on the type of defect, CDG are divided into the following groups: disorders of N-glycosylation or O-glycosylation of proteins, defects in modification of proteins by GPI anchor, disorders of lipid glycosylation and defects that impact multiple glycosylation pathways. The aim of the thesis was to find new biochemical analyses suitable for diagnostics and characterization of CDG patients. The experimental conditions were optimized for selected markers (Sambucus Nigra (SNA) lectin, proaerolysin (FLAER), antibodies to proteins CD55 and CD59) and the staining was applied to cultivated skin fibroblasts from controls and patients diagnosed with CDG by whole-exome sequencing (ATP6AP1-CDG, PIGN-CDG, SLC10A7-CDG, PISD deficiency). The experiments were performed using flow cytometry (FACS) and fluorescent microscopy (FM). The detection of sialylation by SNA lectin and analysis of the mitochondrial membrane potential changes by a fluorescent labelled probe JC-1 with FCCP simulation of mitochondrial...

This diploma thesis deals with the study of film formation in compliant contact. Primary attention is dedicated to the influence of the entrainment speed and slide-to-roll ratio (SRR) on the central film thickness. Based on a research study of the available experimental methods, the fluorescence microscopy was used to achieve the aim of the work. Experiments were realized in ball-on-disk configuration. Firstly, a validation of the method was performed where the solid contact of the ceramic ball and the glass disk was analyzed. Since the measured values were in a good agreement with the theoretical prediction, a ball of NBR rubber material, which is commonly used for production of lip seals and o-rings, was subsequently used. The experimental measurements were performed under pure rolling and rolling/sliding conditions when the entrainment speeds varied in the range from 10 to 400 mm / s. A constant load of 0.2 N was applied. The measured data were then compared to the theoretical predictions for compliant contacts. The results showed that the thickness of the lubricating film gradually increases with increasing entrainment speed, which corresponds to theoretical assumption. Against expectations, evident influence of SRR on the film formation has been observed. In the last part of the thesis the measured data are confronted with previously published studies. Finally, some limitations of this study are discussed and recommendations for further improvement of the used experimental method are suggested.